LOCUS       sequence-342844-        3834 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  This gene was cloned into the gateway middle entry vector. It can be
            used for generating antisense riboprobe or gene expression using a 
            gateway system..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3834)
  AUTHORS   Silic MR, Murata SH, Park SJ, Zhang G
  TITLE     Evolution of inwardly rectifying potassium channels and their gene 
            expression in zebrafish embryos.
  JOURNAL   Dev Dyn. 2021 Sep 23. doi: 10.1002/dvdy.425.
  PUBMED    34558132
REFERENCE   2  (bases 1 to 3834)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Dev Dyn. 
            2021 Sep 23. doi: 10.1002/dvdy.425."
COMMENT     Sequence Label: pDONR221-kcnj2a
FEATURES             Location/Qualifiers
     source          1..3834
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     RBS             1820..1831
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     protein_bind    complement(1959..2058)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2075..2094)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2076..2094)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2099..2115)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2099..2115)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2217..2236)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2228..3037
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(2295..2314)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3184..3772
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3673..3692
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcaggcttc accatgggaa gtgtgcgggc caaccgctac agcattgtgt catcagagga
      721 ggacggtatg aagttggcca ctgctgcggt gccaaatggg tacggtaacg ggaagagcaa
      781 agtgcacacc cgtcaccaaa cccaaagcag gttcgtcaag aaagacggac actgcaacgt
      841 gcagttcatc aatgtcagcg aaaagagcca gcgctacctg gctgacatct tcaccacatg
      901 cgtggacatc cgctggcgat ggatgttcgt tatcttttgc ttggccttcc tgctgtcgtg
      961 gctgtttttt ggatgtatct tctggttggt tgccatattc catggagacc tggagaacaa
     1021 tggtcacaag tgtgtctcca acgtcagtag cttcactgct gcctttctct tctccatcga
     1081 gacgcagact acaatcggct atggttatcg atatgttaca gatgagtgcc ccatcgcggt
     1141 gttcatggtg gtgtttcaaa gcatagttgg ctgcatcatt gacgccttca tcatcggtgc
     1201 agtcatggca aagatggcaa agcccaagaa aaggaacgag acgctggtgt tcagccacaa
     1261 cgctacggtc gccatgaggg acaataagct gtgtctgatg tggagggtgg ggaacttgcg
     1321 caaaagccac ctggtggagg cccatgtccg tgctcagctc ctcaggtctc gcaccacagc
     1381 cgaaggagag tttatccctc ttgaccagat ggacattgat gtaggcttcg acagtggcat
     1441 tgaccgcatc ttcttggtgt cacccatcac catcgtccat gaaatcgacg aggacagccc
     1501 tttctatgac atgagcaaac aagaaatgga aaactctgac ttcgaaattg tggtcatcct
     1561 ggagggcatg gtggaggcta cggcaatgac cacccagtgt cgtagctcct atgtggcgaa
     1621 tgaaatcctc tgggcacacc gctttgagcc agtcctcttt gaggagaaaa actattacaa
     1681 agtagactac tctcgctttc acaagaccta cgaagtcccc agcaccccgc tgtgcagtgc
     1741 aagggacctt gctgagaaaa agtacatcat atccaactct aattcatttt gctatgagaa
     1801 cgaggtggcc ctttcaaaca aagaggagaa agaagaaggg cacggggaca gcctggggcc
     1861 tgggggaaca aacacggaaa ctagctcaga ctctgaacac agccaggcca ccgttccctt
     1921 ggaacctcgg cctctacggc gggaatctga aatatgagac ccagctttct tgtacaaagt
     1981 tggcattata agaaagcatt gcttatcaat ttgttgcaac gaacaggtca ctatcagtca
     2041 aaataaaatc attatttgcc atccagctga tatcccctat agtgagtcgt attacatggt
     2101 catagctgtt tcctggcagc tctggcccgt gtctcaaaat ctctgatgtt acattgcaca
     2161 agataaaaat atatcatcat gaacaataaa actgtctgct tacataaaca gtaatacaag
     2221 gggtgttatg agccatattc aacgggaaac gtcgaggccg cgattaaatt ccaacatgga
     2281 tgctgattta tatgggtata aatgggctcg cgataatgtc gggcaatcag gtgcgacaat
     2341 ctatcgcttg tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag
     2401 cgttgccaat gatgttacag atgagatggt cagactaaac tggctgacgg aatttatgcc
     2461 tcttccgacc atcaagcatt ttatccgtac tcctgatgat gcatggttac tcaccactgc
     2521 gatccccgga aaaacagcat tccaggtatt agaagaatat cctgattcag gtgaaaatat
     2581 tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg attcctgttt gtaattgtcc
     2641 ttttaacagc gatcgcgtat ttcgtctcgc tcaggcgcaa tcacgaatga ataacggttt
     2701 ggttgatgcg agtgattttg atgacgagcg taatggctgg cctgttgaac aagtctggaa
     2761 agaaatgcat aaacttttgc cattctcacc ggattcagtc gtcactcatg gtgatttctc
     2821 acttgataac cttatttttg acgaggggaa attaataggt tgtattgatg ttggacgagt
     2881 cggaatcgca gaccgatacc aggatcttgc catcctatgg aactgcctcg gtgagttttc
     2941 tccttcatta cagaaacggc tttttcaaaa atatggtatt gataatcctg atatgaataa
     3001 attgcagttt catttgatgc tcgatgagtt tttctaatca gaattggtta attggttgta
     3061 acactggcag agcattacgc tgacttgacg ggacggcgca agctcatgac caaaatccct
     3121 taacgtgagt tacgcgtcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     3181 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     3241 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     3301 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     3361 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     3421 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     3481 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     3541 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     3601 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     3661 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     3721 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     3781 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgtt
//
